Interaction Details


Details of experiments between Q04917 and P30305
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPVR
Construct: IRRFQ(pSer)MPVR (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 7.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.01
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (crude synthetic peptide)
Average holdup BI: 0.86
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 130
P value: 8.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.69
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 3

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RNITNSQAPD
Construct: RNITN(pSer)QAPD (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 0.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 4

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: KPTHPSSTHA
Construct: KPTHP(pSer)STHA (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 130
P value: 1.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.43
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 5

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 225-234
Site: 14-3-3_pSer230
Modification: (SEP)230
Sequence: AQRPSSAPDL
Construct: AQRPS(pSer)APDL (crude synthetic peptide)
Average holdup BI: 0.9
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 8.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.86
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 6

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 286-295
Site: 14-3-3_pSer291
Modification: (SEP)291
Sequence: PPGMESLISA
Construct: PPGME(pSer)LISA (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 130
P value: 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 7

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 318-327
Site: 14-3-3_pSer323
Modification: (SEP)323
Sequence: LFRSPSMPCS
Construct: LFRSP(pSer)MPCS (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 130
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 8

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 370-379
Site: 14-3-3_pSer375
Modification: (SEP)375
Sequence: VLRSKSLCHD
Construct: VLRSK(pSer)LCHD (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.42
Immobilized partner concentration (10-6M): 130
P value: 0.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 9

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 460-469
Site: 14-3-3_pSer465
Modification: (SEP)465
Sequence: ERDAESFLLK
Construct: ERDAE(pSer)FLLK (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
P value: 1.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 10

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 558-567
Site: 14-3-3_pSer563
Modification: (SEP)563
Sequence: RLKTRSWAGE
Construct: RLKTR(pSer)WAGE (crude synthetic peptide)
Average holdup BI: 0.86
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 7.39
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.66
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 11

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 122-131
Site: 14-3-3_pThr127
Modification: (TPO)127
Sequence: HMAEQTFEQA
Construct: HMAEQ(pThr)FEQA (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 130
P value: 0.18
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 12

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 162-171
Site: 14-3-3_pThr167
Modification: (TPO)167
Sequence: VLRNITNSQA
Construct: VLRNI(pThr)NSQA (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 0.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 13

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: AFLLQTVDGK
Construct: AFLLQ(pThr)VDGK (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
P value: 1.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 14

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 48-57
Site: 14-3-3_pThr53
Modification: (TPO)53
Sequence: ASSPVTTLTQ
Construct: ASSPV(pThr)TLTQ (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 2.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 15

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 49-58
Site: 14-3-3_pThr54
Modification: (TPO)54
Sequence: SSPVTTLTQT
Construct: SSPVT(pThr)LTQT (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 130
P value: 1.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 16

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 51-60
Site: 14-3-3_pThr56
Modification: (TPO)56
Sequence: PVTTLTQTMH
Construct: PVTTL(pThr)QTMH (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 130
P value: 0.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 17

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: CDC25B
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: TTLTQTMHDL
Construct: TTLTQ(pThr)MHDL (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 4.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))